Methylation-induced silencing of maspin contributes to the proliferation of human glioma cells

Xu,Liang; Liu,Hongyuan; Yu,Ju; Wang,Zhongyong; Zhu,Qing; Li,Zongping; Zhong,Qi; Zhang,Shuyu; Qu,Mingqi; Lan,Qing

doi:10.3892/or.2016.4783

Methylation-induced silencing of maspin contributes to the proliferation of human glioma cells

Authors:
- Liang Xu
- Hongyuan Liu
- Ju Yu
- Zhongyong Wang
- Qing Zhu
- Zongping Li
- Qi Zhong
- Shuyu Zhang
- Mingqi Qu
- Qing Lan
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Affiliations: Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China, Department of Neurosurgery, Mianyang Central Hospital, Mianyang, Sichuan 621000, P.R. China, School of Radiation Medicine and Protection and Jiangsu Provincial Key Laboratory of Radiation Medicine and Protection, Medical College of Soochow University, Suzhou, Jiangsu 215123, P.R. China, Department of Neurosurgery, Henan Provincial People's Hospital, Zhengzhou, Henan 450000, P.R. China
Published online on: May 4, 2016 https://doi.org/10.3892/or.2016.4783
Pages: 57-64
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Maspin, a member of the serpin superfamily of serine protease inhibitors, has been reported to be involved in cancer initiation and progression. However, the expression of maspin and its expression regulation in glioma remain unknown. In the present study, we aimed to investigate the function of maspin in glioma cells and its regulatory mechanism. We found that the expression of maspin was silenced in glioma cells and tissues. Although maspin had no effect on the migration and invasion of human glioma cells in vitro, overexpression of maspin inhibited cell growth in U87 cells. We showed that the methylase inhibitor 5-Aza-2'-deoxycytidine induced the expression of maspin in glioma cell lines. Furthermore, both U87 and U251 cells showed hypermethylation in the maspin promoter. In addition, bisulphite sequencing analysis indicated that 16 CpG sites in the promoter were completely methylated in glioma cells and cancerous tissues, while CpG dinucleotides in the maspin promoter were unmethylated in normal brain tissues. Our data suggest that methylation-induced silencing of maspin contributes to the proliferation of human glioma cells, and maspin may be a potential therapeutic target in glioma.

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1	Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW and Kleihues P: The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 114:97–109. 2007. View Article : Google Scholar : PubMed/NCBI
2	Huse JT, Holland E and DeAngelis LM: Glioblastoma: Molecular analysis and clinical implications. Annu Rev Med. 64:59–70. 2013. View Article : Google Scholar
3	Cloughesy TF, Cavenee WK and Mischel PS: Glioblastoma: From molecular pathology to targeted treatment. Annu Rev Pathol. 9:1–25. 2014. View Article : Google Scholar
4	Burgess DJ: Epigenetics. Dissecting driving DNA methylations. Nat Rev Cancer. 12:448–449. 2012. View Article : Google Scholar : PubMed/NCBI
5	Cahill N and Rosenquist R: Uncovering the DNA methylome in chronic lymphocytic leukemia. Epigenetics. 8:138–148. 2013. View Article : Google Scholar : PubMed/NCBI
6	Sulaiman L, Juhlin CC, Nilsson IL, Fotouhi O, Larsson C and Hashemi J: Global and gene-specific promoter methylation analysis in primary hyperparathyroidism. Epigenetics. 8:646–655. 2013. View Article : Google Scholar : PubMed/NCBI
7	Mueller S, Phillips J, Onar-Thomas A, Romero E, Zheng S, Wiencke JK, McBride SM, Cowdrey C, Prados MD, Weiss WA, et al: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome. Neuro-oncol. 14:1146–1152. 2012. View Article : Google Scholar : PubMed/NCBI
8	Zou Z, Anisowicz A, Hendrix MJ, Thor A, Neveu M, Sheng S, Rafidi K, Seftor E and Sager R: Maspin, a serpin with tumor-suppressing activity in human mammary epithelial cells. Science. 263:526–529. 1994. View Article : Google Scholar : PubMed/NCBI
9	Sharma G, Mirza S, Parshad R, Srivastava A, Gupta SD, Pandya P and Ralhan R: Clinical significance of Maspin promoter methylation and loss of its protein expression in invasive ductal breast carcinoma: Correlation with VEGF-A and MTA1 expression. Tumour Biol. 32:23–32. 2011. View Article : Google Scholar
10	McKenzie S, Sakamoto S and Kyprianou N: Maspin modulates prostate cancer cell apoptotic and angiogenic response to hypoxia via targeting AKT. Oncogene. 27:7171–7179. 2008. View Article : Google Scholar : PubMed/NCBI
11	Yoshizawa K, Nozaki S, Kitahara H, Kato K, Noguchi N, Kawashiri S and Yamamoto E: Expression of urokinase-type plasminogen activator/urokinase-type plasminogen activator receptor and maspin in oral squamous cell carcinoma: Association with mode of invasion and clinicopathological factors. Oncol Rep. 26:1555–1560. 2011.PubMed/NCBI
12	Bodenstine TM, Seftor RE, Khalkhali-Ellis Z, Seftor EA, Pemberton PA and Hendrix MJ: Maspin: Molecular mechanisms and therapeutic implications. Cancer Metastasis Rev. 31:529–551. 2012. View Article : Google Scholar : PubMed/NCBI
13	Wu Y, Alvarez M, Slamon DJ, Koeffler P and Vadgama JV: Caspase 8 and maspin are downregulated in breast cancer cells due to CpG site promoter methylation. BMC Cancer. 10(32)2010. View Article : Google Scholar
14	Alvarez Secord A, Darcy KM, Hutson A, Huang Z, Lee PS, Jewell EL, Havrilesky LJ, Markman M, Muggia F and Murphy SK: The regulation of MASPIN expression in epithelial ovarian cancer: association with p53 status, and MASPIN promoter methylation: a gynecologic oncology group study. Gynecol Oncol. 123:314–319. 2011. View Article : Google Scholar : PubMed/NCBI
15	Rose SL, Fitzgerald MP, White NO, Hitchler MJ, Futscher BW, De Geest K and Domann FE: Epigenetic regulation of maspin expression in human ovarian carcinoma cells. Gynecol Oncol. 102:319–324. 2006. View Article : Google Scholar : PubMed/NCBI
16	Zhang W and Zhang M: Tissue microarray analysis of maspin expression and its reverse correlation with mutant p53 in various tumors. Int J Oncol. 20:1145–1150. 2002.PubMed/NCBI
17	Xie R, Yang H, Xiao Q, Mao F, Zhang S, Ye F, Wan F, Wang B, Lei T and Guo D: Downregulation of LRIG1 expression by RNA interference promotes the aggressive properties of glioma cells via EGFR/Akt/c-Myc activation. Oncol Rep. 29:177–184. 2013.
18	Squatrito M, Brennan CW, Helmy K, Huse JT, Petrini JH and Holland EC: Loss of ATM/Chk2/p53 pathway components accelerates tumor development and contributes to radiation resistance in gliomas. Cancer Cell. 18:619–629. 2010. View Article : Google Scholar : PubMed/NCBI
19	Chow LM, Endersby R, Zhu X, Rankin S, Qu C, Zhang J, Broniscer A, Ellison DW and Baker SJ: Cooperativity within and among Pten, p53, and Rb pathways induces high-grade astrocytoma in adult brain. Cancer Cell. 19:305–316. 2011. View Article : Google Scholar : PubMed/NCBI
20	Stark AM, Schem C, Maass N, Hugo HH, Jonat W, Mehdorn HM and Held-Feindt J: Expression of metastasis suppressor gene maspin is reduced in breast cancer brain metastases and correlates with the estrogen receptor status. Neurol Res. 32:303–308. 2010. View Article : Google Scholar
21	Dzinic SH, Chen K, Thakur A, Kaplun A, Bonfil RD, Li X, Liu J, Bernardo MM, Saliganan A, Back JB, et al: Maspin expression in prostate tumor elicits host anti-tumor immunity. Oncotarget. 5:11225–11236. 2014. View Article : Google Scholar : PubMed/NCBI
22	Prasad CP, Rath G, Mathur S, Bhatnagar D and Ralhan R: Expression analysis of maspin in invasive ductal carcinoma of breast and modulation of its expression by curcumin in breast cancer cell lines. Chem Biol Interact. 183:455–461. 2010. View Article : Google Scholar
23	Machtens S, Serth J, Bokemeyer C, Bathke W, Minssen A, Kollmannsberger C, Hartmann J, Knüchel R, Kondo M, Jonas U, et al: Expression of the p53 and Maspin protein in primary prostate cancer: Correlation with clinical features. Int J Cancer. 95:337–342. 2001. View Article : Google Scholar : PubMed/NCBI
24	Machowska M, Wachowicz K, Sopel M and Rzepecki R: Nuclear location of tumor suppressor protein maspin inhibits proliferation of breast cancer cells without affecting proliferation of normal epithelial cells. BMC Cancer. 14:1422014. View Article : Google Scholar : PubMed/NCBI
25	Cher ML, Biliran HR Jr, Bhagat S, Meng Y, Che M, Lockett J, Abrams J, Fridman R, Zachareas M and Sheng S: Maspin expression inhibits osteolysis, tumor growth, and angiogenesis in a model of prostate cancer bone metastasis. Proc Natl Acad Sci USA. 100:7847–7852. 2003. View Article : Google Scholar : PubMed/NCBI
26	Bernardo MM, Meng Y, Lockett J, Dyson G, Dombkowski A, Kaplun A, Li X, Yin S, Dzinic S, Olive M, et al: Maspin reprograms the gene expression profile of prostate carcinoma cells for differentiation. Genes Cancer. 2:1009–1022. 2011. View Article : Google Scholar
27	Kaplun A, Dzinic S, Bernardo M and Sheng S: Tumor suppressor maspin as a rheostat in HDAC regulation to achieve the fine-tuning of epithelial homeostasis. Crit Rev Eukaryot Gene Expr. 22:249–258. 2012. View Article : Google Scholar : PubMed/NCBI
28	Li X, Yin S, Meng Y, Sakr W and Sheng S: Endogenous inhibition of histone deacetylase 1 by tumor-suppressive maspin. Cancer Res. 66:9323–9329. 2006. View Article : Google Scholar : PubMed/NCBI
29	Yin S, Lockett J, Meng Y, Biliran H Jr, Blouse GE, Li X, Reddy N, Zhao Z, Lin X, Anagli J, et al: Maspin retards cell detachment via a novel interaction with the urokinase-type plasminogen activator/urokinase-type plasminogen activator receptor system. Cancer Res. 66:4173–4181. 2006. View Article : Google Scholar : PubMed/NCBI
30	Bailey CM, Khalkhali-Ellis Z, Kondo S, Margaryan NV, Seftor RE, Wheaton WW, Amir S, Pins MR, Schutte BC and Hendrix MJ: Mammary serine protease inhibitor (Maspin) binds directly to interferon regulatory factor 6: Identification of a novel serpin partnership. J Biol Chem. 280:34210–34217. 2005. View Article : Google Scholar : PubMed/NCBI
31	Endsley MP, Hu Y, Deng Y, He X, Warejcka DJ, Twining SS, Gonias SL and Zhang M: Maspin, the molecular bridge between the plasminogen activator system and beta1 integrin that facilitates cell adhesion. J Biol Chem. 286:24599–24607. 2011. View Article : Google Scholar : PubMed/NCBI
32	Blacque OE and Worrall DM: Evidence for a direct interaction between the tumor suppressor serpin, maspin, and types I and III collagen. J Biol Chem. 277:10783–10788. 2002. View Article : Google Scholar : PubMed/NCBI
33	Li X, Kaplun A, Lonardo F, Heath E, Sarkar FH, Irish J, Sakr W and Sheng S: HDAC1 inhibition by maspin abrogates epigenetic silencing of glutathione S-transferase pi in prostate carcinoma cells. Mol Cancer Res. 9:733–745. 2011. View Article : Google Scholar : PubMed/NCBI
34	Latha K, Zhang W, Cella N, Shi HY and Zhang M: Maspin mediates increased tumor cell apoptosis upon induction of the mitochondrial permeability transition. Mol Cell Biol. 25:1737–1748. 2005. View Article : Google Scholar : PubMed/NCBI
35	Toillon RA, Lagadec C, Page A, Chopin V, Sautière PE, Ricort JM, Lemoine J, Zhang M, Hondermarck H and Le Bourhis X: Proteomics demonstration that normal breast epithelial cells can induce apoptosis of breast cancer cells through insulin-like growth factor-binding protein-3 and maspin. Mol Cell Proteomics. 6:1239–1247. 2007. View Article : Google Scholar : PubMed/NCBI
36	Zhang S, Hao J, Xie F, Hu X, Liu C, Tong J, Zhou J, Wu J and Shao C: Downregulation of miR-132 by promoter methylation contributes to pancreatic cancer development. Carcinogenesis. 32:1183–1189. 2011. View Article : Google Scholar : PubMed/NCBI
37	Sarkar S, Goldgar S, Byler S, Rosenthal S and Heerboth S: Demethylation and re-expression of epigenetically silenced tumor suppressor genes: Sensitization of cancer cells by combination therapy. Epigenomics. 5:87–94. 2013. View Article : Google Scholar : PubMed/NCBI
38	Jaenisch R and Bird A: Epigenetic regulation of gene expression: How the genome integrates intrinsic and environmental signals. Nat Genet. 33(Suppl): 245–254. 2003. View Article : Google Scholar : PubMed/NCBI
39	Maass N, Biallek M, Rösel F, Schem C, Ohike N, Zhang M, Jonat W and Nagasaki K: Hypermethylation and histone deacetylation lead to silencing of the maspin gene in human breast cancer. Biochem Biophys Res Commun. 297:125–128. 2002. View Article : Google Scholar : PubMed/NCBI
40	Reddy GP, Barrack ER, Dou QP, Menon M, Pelley R, Sarkar FH and Sheng S: Regulatory processes affecting androgen receptor expression, stability, and function: Potential targets to treat hormone-refractory prostate cancer. J Cell Biochem. 98:1408–1423. 2006. View Article : Google Scholar : PubMed/NCBI
41	Eitel JA, Bijangi-Vishehsaraei K, Saadatzadeh MR, Bhavsar JR, Murphy MP, Pollok KE and Mayo LD: PTEN and p53 are required for hypoxia induced expression of maspin in glioblastoma cells. Cell Cycle. 8:896–901. 2009. View Article : Google Scholar : PubMed/NCBI
42	Jiang R, Xia Y, Li J, Deng L, Zhao L, Shi J, Wang X and Sun B: High expression levels of IKKalpha and IKKbeta are necessary for the malignant properties of liver cancer. Int J Cancer. 126:1263–1274. 2010. View Article : Google Scholar
43	Zhang M: PTEN in action: Coordinating with p53 to regulate maspin gene expression. Cell Cycle. 8:1112–1113. 2009. View Article : Google Scholar : PubMed/NCBI